Sequential ChIP / ChIP-reChIP - Which of the two antibodies to use first? (Nov/17/2010 )
I've just started doing ChIP-reChIP experiments using antibodies against histone modifications to establish the method in our lab.
My question now is whether there is any consensus on which of the two antibodies to use in the first round of ChIP, i.e. should I do the first
ChIP against the more abundant protein and then the second round against the less abundant protein as one would assume intuitively, or is it the opposite way?
Thanks on any thoughts/comments,
Alright, found what I needed. If anyone should have similar questions, for future reference:
Curr Protoc Mol Biol. 2005 May;Chapter 21:Unit 21.8.
Analysis of protein co-occupancy by quantitative sequential chromatin immunoprecipitation.
Geisberg JV, Struhl K.